ES2724199T3 - Vehicle for adsorption of blood components and column for adsorption of blood components - Google Patents

Abstract

Vehicle for adsorption of blood components comprising a water insoluble vehicle composed of a fiber or particle, said water insoluble vehicle having a surface on which one or more functional groups are introduced, said one or more functional groups containing a functional group acid selected from the group consisting of the sulfate group, sulfite group and sulfonate group; and containing an amino group, said fiber having a fiber diameter of or having said particle having a particle diameter of 0.5 to 20 μm, and wherein said water insoluble vehicle has a negative charge amount of 1, 0x10-4 to 1,5x10-3 eq / g.

Description

DESCRIPTION

Vehicle for adsorption of blood components and column for adsorption of blood components TECHNICAL SECTOR

The present invention relates to a vehicle for adsorption of blood components and a column for adsorption of blood components.

TECHNICAL BACKGROUND

Humoral factors, such as inflammatory cytokines, are deeply implicated in the causes of inflammatory diseases, such as systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, ulcerative colitis and Crohn's disease, and attempts are being made to treat inflammatory diseases by the inactivation of these humoral factors with biological products such as drugs and low molecular weight antibodies. However, each of these humoral factors does not act only at the site of inflammation, but multiple humoral factors act synergistically to cause the development and progression of inflammatory diseases. Therefore, recent interest has focused on leukocytopheresis in which activated leukocytes are removed per se as a source of humoral factors.

Leukocitapheresis is a therapeutic procedure by purification of blood in which blood is drawn from the vein and activated leukocytes are removed from it using, for example, a column filled with a vehicle for adsorption, followed by returning blood. purified to the vein on the opposite side. Leukocytes can be divided into approximately 3 types, that is, granulocytes, monocytes and lymphocytes. Since leukocytes directly involved in inflammation caused by inflammatory diseases are considered granulocytes and monocytes, a vehicle that selectively adsorbs granulocytes (patent document 1) and a vehicle that selectively adsorbs both granulocytes and activated monocytes as inflammatory cytokines have been developed ( patent documents 2 and 3).

On the other hand, it is also known that lymphocytes are indirectly involved in inflammation caused by inflammatory diseases, and there is also the opinion that not only granulocytes and monocytes, but also lymphocytes must be removed by adsorption to achieve rapid cure of a serious inflammatory disease in a patient.

Therefore, a vehicle has also been developed that can adsorb all 3 types of leukocytes, that is, granulocytes, monocytes and lymphocytes (patent documents 4 and 5). EP561379 discloses a filter means for treating a blood material comprising a porous polymeric element that has, on a surface portion thereof, a negative charge and that has a surface electric charge in the range of -1x10-9 a -3x10-5 eq / g of the polymeric porous element.

DOCUMENTS OF THE PREVIOUS TECHNIQUE

[Patent documents]

[Patent Document 1] JP 2501500 B

[Patent document 2] JP 2006-312804 A

[Patent Document 3] JP 7-080062 A

[Patent Document 4] JP 60-193468 A

[Patent Document 5] JP 2001-310917 A

CHARACTERISTICS OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

However, the vehicle that allows the adsorption of the 3 types of leukocytes, that is, granulocytes, monocytes and lymphocytes, is composed of a nonwoven fabric prepared from an ultrafine fiber, a nonwoven fabric having a high apparent density. Since most of the blood components other than erythrocytes are filtered and pressure loss occurs during blood circulation, safety problems such as stopping circulation and blood leakage have been reported. On the other hand, although a powerful leukocyte removal vehicle has been demanded that can adsorb not only the 3 types of leukocytes but also the inflammatory cytokines, so far a successful development of said vehicle has not been reported due to the difficulty to Overcome the previous problem.

In view of this, the present invention is intended to disclose a vehicle for adsorption of blood components, which can eliminate leukocytes, including all granulocytes, monocytes and lymphocytes by adsorption, while suppressing pressure loss during circulation blood, a vehicle that can also eliminate inflammatory cytokines by adsorption at the same time.

MEANS TO SOLVE PROBLEMS

In order to solve the problem described above, the inventors of the present invention studied intensively to discover a vehicle for adsorption of blood components that causes less pressure loss due to obstruction and can eliminate granulocytes, monocytes and lymphocytes, as well as cytokines. Inflammatory with high adsorption efficiency, thus completing the present invention.

That is, the present invention discloses the vehicle for adsorption of blood components and the column for adsorption of blood components described in (1) to (6) below.

(1) A vehicle for the adsorption of blood components comprising a water insoluble vehicle composed of a fiber or particle, the water insoluble vehicle having a surface on which one or more functional groups are introduced, containing the functional group (s) an acid functional group selected from the group consisting of the sulfate group, sulphite group and sulfonate group; and which contains an amino group; the fiber having a fiber diameter or the particle having a particle diameter of 0.5 to 20 pm and wherein said water insoluble vehicle has a negative charge amount of 1.0 x 10-4 to 1.5 x 10-3 eq / g.

(2) The vehicle for adsorption of blood components, according to (1), in which the water insoluble vehicle has a porosity of 85 to 98%.

(3) The vehicle for adsorption of blood components, according to any of (1) to (2), in which the water insoluble vehicle is a fiber having a fiber diameter of 4 to 10 pm.

(4) The vehicle for adsorption of blood components, according to any one of (1) to (3), in which the acid functional group and the amino group are linked to each other through an alkyl chain.

(5) The vehicle for adsorption of blood components, according to (4), in which the alkyl chain is an alkyl chain having no more than 3 carbon atoms.

(6) A column for adsorption of blood components filled with the vehicle for adsorption of blood components, according to any of claims (1) to (5).

EFFECT OF THE INVENTION

With the vehicle for adsorption of the blood components of the present invention, the 3 types of leukocytes, i.e. granulocytes, monocytes and lymphocytes, can be efficiently removed by adsorption of blood from a patient with an inflammatory disease, and the Inflammatory cytokines can also be removed by adsorption at the same time. In addition, a column for adsorption of blood components filled with the vehicle for adsorption of blood components of the present invention can be used for leukocytopheresis and can be suitably used in the treatment of a serious inflammatory disease. BETTER WAY TO CARRY OUT THE INVENTION

The vehicle for adsorption of the blood components of the present invention comprises a water insoluble vehicle composed of a fiber or particle, a water insoluble vehicle having a surface in which one or more functional groups are introduced, in which the group or functional groups contain a functional acid group selected from the group consisting of the sulfate group, sulfite group and sulfonate group; and contain an amino group, fiber having a fiber diameter or a particle having a particle diameter of 0.5 to 20 pm and in which said water insoluble vehicle has a negative charge amount of 1.0 x 10 -4 to 1.5 x 10-3 eq / g. The "vehicle for adsorption of blood components" means a material with which one or more blood components can be removed from the blood by adsorption.

The blood component means a component that constitutes blood, and examples of blood components include blood cell components such as erythrocytes, leukocytes and platelets; and humoral factors such as inflammatory cytokines. For the purpose of treating an inflammatory disease, leukocytes and inflammatory cytokines are preferably removed by adsorption.

Inflammatory cytokine means a protein that is secreted from a cell and transmits information to a specific cell. Examples of inflammatory cytokine include interleukins, tumor necrosis factor-a, factor transforming growth beta, interferon-y (hereinafter referred to as INF-y), angiogenic growth factors and immunosuppressive acid protein.

Interleukin means a cytokine that is secreted from a leukocyte and works to control the immune system, and examples of interleukin include interleukin-1, interleukin-6, interleukin-8 (hereinafter referred to as IL-8), interleukin -10 and interleukin-17 (hereinafter referred to as IL-17).

Adsorption means a state in which a blood component adsorbs to the vehicle for adsorption of blood components and does not separate easily.

Examples of the "water insoluble carrier compound of a fiber or particle" material include polyolefins such as polyethylene and polypropylene; polyesters such as polyethylene terephthalate and polybutylene terephthalate; fluorinated polymers such as Teflon (registered trademark); polysulfone-based polymers such as poly (p-phenylene ether sulfone); polyetherimides; polyimides; polyamides; polyethers; polyphenylene sulfides; polystyrenes; and acrylic polymers; and materials prepared by mixing or alloying these macromolecular compounds. For easy introduction of a functional group to the surface of the water-insoluble vehicle, polystyrenes are preferred, and, in view of heat resistance or shape retention during processing, polypropylene or polypropylene-polyethylene copolymers are preferred .

The "functional group containing an acid functional group selected from the group consisting of the sulfate group, the sulphite group and the sulfonate group; and which contains an amino group" means a functional group that contains, in a part of the chemical structure of the functional group, at least one of each: an acid functional group selected from the group consisting of the sulfate group, the sulphite group and the sulphonate group; and an amino group. The sulfate group (-OSO2OH), sulphite group (-O (SO) OH) and sulfonate group (-SO2OH) have similar chemical structures to each other and each of these groups has an acidic hydroxyl group. Therefore, these groups show common characteristics such as strong acidity and negativity.

The acid functional group is preferably placed at one end of the functional group described above with a view to achieving easy interactions between the acid functional group and the blood components.

The amino group is preferably a secondary amino group, more preferably, a tertiary amino group. In the chemical structure of the functional group described above, the chemical structure existing between the acidic functional group and the amino group, that is, the chemical structure that links the acidic functional group to the amino group (hereinafter referred to as the spacer), is preferably consisting of one or more hydrogen atoms, carbon atoms, oxygen atoms, nitrogen atoms, sulfur atoms and / or silicon atoms. The spacer is, more preferably, an alkyl chain, even more preferably an alkyl chain having no more than 3 carbon atoms. In cases where the spacer is too large, the density of the acid functional group decreases, so that the number of atoms constituting the spacer is preferably no more than 200.

Examples of the reactive functional group that mediates the binding between the water insoluble vehicle and the functional group described above when the "functional group containing an acid functional group selected from the group consisting of the sulfate group, sulphite group and sulfonate group; and which contains an amino group "must be introduced into the surface of the water insoluble vehicle includes active halogen groups such as the halomethyl group, haloacetyl group, haloacetamidomethyl group and halogenated alkyl group; epoxy group; carboxyl group; isocyanate group; thioisocyanate group; and acid anhydride group. To have an appropriate degree of reactivity, active halogen groups are preferred, and the haloacetamidomethyl group is more preferred.

The functional group described above in which an acid functional group is positioned as a term and the spacer is an alkyl chain can be obtained, for example, by reacting an aminoalkyl sulfonic acid readily available on the market with a haloacetamidomethyl group. In particular, the functional group described above in which the spacer is an alkyl chain having no more than 3 carbon atoms can be obtained by reacting aminoethylsulfonic acid (hereinafter referred to as taurine) or 3-aminopropiosulfonic acid (hereinafter referred to as homotaurin ), or N-methylaminoethylsulfonic acid (hereinafter referred to as N-methyltaurine) or 1,3-propanosultone (hereinafter referred to as propanosultone) with a haloacetamidomethyl group.

The "fiber diameter of the fiber" and the "particle diameter of the particle" of the "water insoluble vehicle composed of a fiber or particle" must be "0.5 to 20 | um" to perform the phagocytic activity of the leukocytes, and the fiber diameter and the particle diameter are preferably from 4 to 20 µm, more preferably, from 4 to 10 µm in view of a more stable execution of the phagocytic activity. The lower limit of the fiber diameter and the particle diameter is preferably 0.5 µm, more preferably 4 µm. The upper limit of the fiber diameter and the particle diameter is preferably 20 | um, more preferably 10 | um. Any of the preferred lower limits may be combined with any of the preferred upper limits. Activity Phagocytic leukocytes in this document means the property of granulocytes and monocytes to capture and eat microorganisms and bacteria that have invaded the human body or the like.

The "fiber diameter of a fiber" means the average of the values obtained by randomly collecting 10 samples of small fragments of the fiber and taking a photograph of each sample using a scanning electron microscope with a magnification of 2,000 x, followed by the Fiber diameter measurement in 10 locations per photo (100 locations in total). Similarly, the "particle diameter of a particle" means the average of the values obtained by randomly collecting 10 samples of small pieces of the particle and taking a photograph of each sample using a scanning electron microscope with an increase of 2,000 x, followed by the measurement of the particle diameter in 10 locations per photograph (100 locations in total).

In cases where the fiber diameter of the fiber is less than 10 pm, a fiber having a larger diameter may be mixed in order to ensure the strength of the vehicle for adsorption of the blood components, and the diameter of fiber of said fiber having a larger diameter is preferably from 10 to 50 pm.

Examples of the water-insoluble vehicle shape composed of a fiber include a woven fabric, non-woven fabric, cotton fabric and hollow fiber. In cases where the shape is a non-woven fabric, a frame fiber, such as polypropylene, is preferably included to maintain the shape.

The elimination of blood components using the vehicle for adsorption of blood components of the present invention is not based on the principle of filtration, but on the use of the phagocytic capacity of granulocytes and monocytes and on the use of lymphocyte interactions and inflammatory cytokines with the "functional group containing an acid functional group selected from the group consisting of the sulfate group, sulfite group and sulfonate group and containing an amino group", to remove the respective blood components by adsorption. Therefore, in cases where the vehicle for adsorption of the blood components of the present invention is used by filling a container, such as a column with the vehicle, the porosity can be increased and the pressure loss can be greatly reduced by comparison with cases in which a conventional technique is used. On the other hand, since, in cases where the porosity is too large, the shape of the adsorption vehicle can barely be maintained, the porosity of the water insoluble vehicle is preferably 85 to 98%, more preferably 90 to 95% The lower limit of the porosity is preferably 85%, more preferably, 90%. The upper limit of porosity is preferably 98%, more preferably 95%. Any of the preferred lower limits may be combined with any of the preferred upper limits.

"Porosity" is the ratio of the volume of the vacuum in the vehicle for the adsorption of the blood components, and means the percentage value calculated by dividing the volume of the vacuum in the vehicle for the adsorption of the blood components by the apparent volume of the vehicle. for the adsorption of blood components. More specifically, a cross-sectional photograph of the vehicle was taken for adsorption of blood components using a scanning electron microscope with a magnification of 200x, and, according to the result of the image analysis of the photograph, the porosity was calculated according to the Equation 1 next.

Porosity (%) = {(b-a) / bx100 Equation 1

a: The area of the part occupied by the water insoluble vehicle.

b: The total area of the cross section of the vehicle for adsorption of blood components

It is assumed that the acid functional group contained in the "functional group containing an acid functional group selected from the group consisting of the sulfate group, the sulphite group and the sulfonate group; and which contains an amino group" greatly contributes to the lymphocyte adsorption On the other hand, in cases where the density of the acid functional group is too large, it is assumed that competitive adsorption occurs between lymphocytes and proteins with positive charges, which leads to a reduced rate of lymphocyte adsorption. The density of the acid functional group can be represented as the amount of negative charge. The amount of negative charge of the vehicle for adsorption of the blood components of the present invention is 1.5 x 10'-5eq / g, more preferably 1.0 x 10-4eq / g.

A negative charge amount of 1 eq / g herein means that 1 g of the adsorption vehicle can adsorb 1 mol of protons.

It is also assumed that the acid functional group contained in the "functional group containing an acid functional group selected from the group consisting of the sulfate group, the sulphite group and the sulfonate group; and containing an amino group" contributes to the adsorption of inflammatory cytokines to some extent. That is, since inflammatory cytokines are approximately 1 to several 10 kDa proteins that comprise many types of ionic amino acids, it is assumed that the portions that have positive charges on the protein molecules interact with the negative acid functional group.

The shape of the column container for adsorption of blood components of the present invention filled with the vehicle for adsorption of blood components is not restricted, provided that the container has an inlet and outlet for blood, and among the examples of the container Prism-shaped containers are included, such as cylindrical containers, in the form of a triangular prism, in the form of a quadrangular prism, in the form of a hexagonal prism and in the form of an octagonal prism. The container is preferably a container that can be filled with the vehicle for adsorption of blood components in a laminated form, a container that can be filled with the vehicle for adsorption of blood components wound in a cylindrical form, or a container in the that blood flows from the circumference of a cylinder inside, followed by a flow out of the container.

EXAMPLES

The column for adsorption of blood components of the present invention will be described in more detail below by means of experimental examples. In the examples,% p represents% by weight.

(Preparation of non-woven fabric made of PP)

A fiber composed of marine islands with 36 islands was obtained, each of which also has a core / shell complex using the following components under the conditions of a turning speed of 800 m / minute and a stretch ratio of 3.

The core component of the island: polypropylene.

The envelope component of the island: 90% by weight of polystyrene and 10% by weight of polypropylene.

The marine component: copolymerized polyester comprising ethylene terephthalate units as the main repeating units and 3% by weight of 5-sodium sulfoisophthalic acid as the copolymerization component.

The composite ratio (weight ratio): core: shell: marine = 45:40:15

After preparing a nonwoven fabric composed of this fiber in an amount of 85% by weight and a polypropylene fiber with a diameter of 20 pm in an amount of 15% by weight, a sheet-shaped polypropylene net (thickness, 0.5 mm; single fiber diameter, 0.3 mm; aperture, 2 mm x 2 mm) was placed between two sheets of this nonwoven fabric, and the result was pierced with a needle to obtain a nonwoven fabric that had a three layer structure (hereinafter referred to as the nonwoven fabric made of PP).

(Preparation of non-woven fabric made of PSt PP)

The non-woven fabric made of PP was treated at 95 ° C with a 3% by weight aqueous solution of sodium hydroxide to dissolve the marine component. Therefore, a nonwoven fabric having a core fiber / shell diameter of 5 pm and an apparent density of 0.02 g / cm3 (nonwoven fabric made of PSt PP, hereinafter referred to as nonwoven fabric) TO).

(Preparation of non-woven fabric modified with chloroacetamidomethyl)

At no more than 10 ° C, 46% by weight of nitrobenzene, 46% by weight of sulfuric acid, 1% by weight of paraformaldehyde and 7% by weight of N-methylol-a-chloracetamide (in hereinafter referred to as NMCA), and the resulting mixture was stirred and dissolved to prepare a reaction liquid for NMCA modification. The temperature of this reaction liquid for the NMCA modification was adjusted to 5 ° C, and the reaction liquid for the NMCA modification was added to the non-woven fabric A in a solid / liquid ratio corresponding to approximately 40 ml of the liquid The reaction mixture for NMCA modification with respect to 1 g of the non-woven fabric A. The reaction mixture was allowed to stand at 5 ° C in a water bath to allow the reaction to proceed for 2 hours. Next, the nonwoven fabric was removed from the reaction mixture and immersed in nitrobenzene in the same amount as the reaction liquid for the NMCA treatment for washing. Subsequently, the nonwoven fabric was removed therefrom, and immersed in methanol for washing, to obtain a nonwoven fabric modified with chloroacetamidomethyl (hereinafter referred to as the nonwoven fabric B).

(Preparation of non-woven fabric modified with tetraethylenepentamine)

Tetraethylenepentamine (hereinafter referred to as TEPA) and triethylamine were dissolved in 500 ml of dimethylsulfoxide (hereinafter referred to as "DMSO"), so that their concentrations were 20 mM and 473 mM, respectively. In the resulting solution, 10 g of the nonwoven fabric B was immersed, and the reaction was allowed to proceed at 40 ° C for 3 hours. The non-woven fabric after the reaction was washed with DMSO and methanol, and, in addition with water, to obtain a non-woven fabric modified with TEPA (hereinafter, the non-woven fabric C). The structural formula of the functional group introduced in the nonwoven fabric C is shown in table 1.

(Preparation of non-woven fabric modified with sulfoethanoamino)

To 500 ml of DMSO, 13 g of taurine were added and triethylamine was added at a concentration of 473 mM. In the resulting mixture, 10 g of the nonwoven fabric B was immersed, and the reaction was allowed to proceed at 70 ° C for 6 hours. The nonwoven fabric after the reaction was washed with DMSO and methanol, and, in addition, with water, to obtain a nonwoven fabric modified with sulfoethanoamino (hereinafter referred to as nonwoven fabric D). The structural formula of the functional group introduced in the nonwoven fabric D is shown in table 1.

(Preparation of non-woven fabric modified with sulfopropanoamino)

To 500 ml of DMSO, 13 g of homotaurin was added and triethylamine was added at a concentration of 473 mM. In the resulting mixture, 10 g of the nonwoven fabric B was immersed, and the reaction was allowed to proceed at 70 ° C for 6 hours. The nonwoven fabric after the reaction was washed with DMSO and methanol, and, in addition, with water, to obtain a sulfopropanoamine modified nonwoven fabric (hereinafter referred to as nonwoven fabric E). The structural formula of the functional group introduced in the non-woven fabric E is shown in Table 1.

(Preparation of modified non-woven fabric with methylsulfoetanamine)

To 500 ml of DMSO, 4.2 g of N-methyltaurine and 5 g of potassium iodide were added, and triethylamine was added additionally at a concentration of 473 mM. In the resulting mixture, 10 g of the nonwoven fabric B was immersed and the reaction was allowed to proceed at 60 ° C for 6 hours. The non-woven fabric after the reaction was washed with DMSO and methanol, and, in addition, with water, to obtain a non-woven fabric modified with methylsulfoananamine (hereinafter referred to as the non-woven fabric F). The structural formula of the functional group introduced in the non-woven fabric E is shown in Table 1.

(Preparation of non-woven fabric modified with dimethylamino)

To 500 ml of DMSO, 2.5 g of dimethylamine and 5 g of potassium iodide were added, and triethylamine was added at a concentration of 473 mM. In the resulting mixture, 10 g of the nonwoven fabric B was immersed, and the reaction was allowed to proceed at 50 ° C for 8 hours. The nonwoven fabric after the reaction was washed with DMSO and methanol, and, in addition, with water, to obtain a nonwoven fabric modified with dimethylamino (hereinafter, nonwoven fabric G). The structural formula of the functional group introduced in the non-woven fabric F is shown in Table 1.

(Preparation of dimethylsulfopropanamine modified nonwoven fabric)

To 465 ml of THF, 9.77 ml of propanosultone was added, and the resulting mixture was mixed, followed by immersion of 10 g of the non-woven fabric G therein and allowing the reaction to proceed at 50 ° C for 6 hours. . The nonwoven fabric after the reaction was washed with THF and methanol, and also with water, to obtain a nonwoven fabric modified with dimethylsulfopropanamine (hereinafter referred to as nonwoven fabric H). The structural formula of the functional group introduced in the nonwoven fabric H is shown in table 1.

(Preparation of non-woven fabric modified with mercaptoethanoamino)

To 500 ml of DMSO, 11.6 g of aminoethanothiol hydrochloride was added and triethylamine was added additionally at a concentration of 473 mM. In the resulting mixture, 10 g of the nonwoven fabric B was immersed, and the reaction was allowed to proceed at 70 ° C for 6 hours. The non-woven fabric after the reaction was washed with DMSO and methanol, and, in addition, with water, to obtain a non-woven fabric modified with mercaptoethanoamine (hereinafter, the non-woven fabric I). The structural formula of the functional group introduced in the non-woven fabric I is shown in Table 1.

(Preparation of non-woven fabric modified with dimethylmercaptoethanoamino)

To 500 ml of DMSO, 4.2 g of dimethylaminoethanethiol hydrochloride and 5 g of potassium iodide were added, and triethylamine was added additionally at a concentration of 473 mM. In the resulting mixture, 10 g of the nonwoven fabric B was immersed and the reaction was allowed to proceed at 60 ° C for 6 hours. The nonwoven fabric after the reaction was washed with DMSO and methanol, and also with water, to obtain a nonwoven fabric modified with dimethylmercaptoethanoamino (hereinafter referred to as the nonwoven fabric J). The structural formula of the functional group introduced in the non-woven fabric J is shown in table 1.

(Preparation of polysulfone modified with chloroacetamidomethyl)

To 32 ml of 5% by weight polysulfone / nitrobenzene solution, 2 ml of 2% by weight NMCA / sulfuric acid solution prepared at 0 ° C was added, and the resulting mixture was stirred for 1 hour. To this mixture, 800 ml of ice cold methanol was added to precipitate the chloroacetamidomethyl modified polysulfone, which was then recovered. The recovered chloroacetamidomethyl polysulfone was dissolved in 20 ml of dimethylformamide (hereinafter referred to as DMF), and 400 ml of ice cold methanol was added back to the resulting solution to obtain polysulfone modified with chloroacetamidomethyl.

(Preparation of non-woven polysulfone fabric modified with tetraethylenepentamine)

In 30 ml of DMF, 1 g of the polysulfone modified with chloroacetamidomethyl was dissolved and tetraethylenepentamine was added to the resulting solution at a concentration of 20 mM. The resulting mixture was stirred for 17 hours and 600 ml of ice cold methanol was added to precipitate the tetraethylenepentamine modified polysulfone, which was then recovered. The polysulfone modified with recovered tetraethylenepentamine was dissolved again in 20 ml of DMF and 0.1 g of the non-woven fabric A was immersed in the resulting solution. The non-woven fabric A was immediately removed from the solution, and was further immersed in methanol, to obtain a non-woven polysulfone fabric modified with tetraethylenepentamine (hereinafter referred to as the non-woven fabric K). The structural formula of the functional group introduced in the non-woven fabric I is shown in Table 1.

(Preparation of polysulfone nonwoven fabric modified with sulfoethanoamino)

In 30 ml of DMF, 1 g of the chloroacetamidomethyl modified polysulfone was dissolved, and taurine was added to the resulting solution at a concentration of 200 mM. The resulting mixture was stirred for 17 hours and 600 ml of ice cold methanol was added to precipitate the sulfoethanoamine modified polysulfone, which was then recovered. The polysulfone modified with sulfoethanoamino recovered was again dissolved in 20 ml of DMF, and 0.1 g of the nonwoven fabric A was immersed in the resulting solution. The non-woven fabric A was immediately removed from the solution, and was further immersed in methanol, to obtain a polysulfone fabric modified with sulfoethanoamino (hereinafter referred to as non-woven fabric L). The structural formula of the functional group introduced in the non-woven fabric L is shown in table 1.

(Preparation of polysulfone non-woven fabric modified with sulfopropanoamine)

In 30 ml of DMF, 1 g of the chloroacetamidomethyl modified polysulfone was dissolved and homotaurin was added to the resulting solution at a concentration of 200 mM. The resulting mixture was stirred for 17 hours and 600 ml of ice cold methanol was added to precipitate the sulfopropanoamine modified polysulfone, which was then recovered. The recovered sulfopropanoamine modified polysulfone was dissolved again in 20 ml of DMF, and 0.1 g of the nonwoven fabric A was immersed in the resulting solution. The non-woven fabric A was immediately removed from the solution, and further immersed in methanol, to obtain a polysulfone non-woven fabric modified with sulfopropanoamino (hereinafter referred to as non-woven fabric M). The structural formula of the functional group introduced in the non-woven fabric M is shown in table 1.

(Preparation of non-woven polysulfone fabric modified with mercaptoethanoamino)

In 30 ml of DMF, 1 g of the chloroacetamidomethyl modified polysulfone was dissolved and aminoethanethiol hydrochloride was added to the resulting solution at a concentration of 200 mM. The resulting mixture was stirred for 17 hours and 600 ml of ice cold methanol was added to precipitate the mercaptoethanoamine-modified polysulfone, which was then recovered. The recovered mercaptoethanoamino modified polysulfone was dissolved again in 20 ml of DMF, and 0.1 g of the nonwoven fabric A was immersed in the resulting solution. The non-woven fabric A was immediately removed from the solution, and further immersed in methanol, to obtain a non-woven fabric of polysulfone modified with mercaptoethanoamine (hereinafter referred to as the non-woven fabric N). The structural formula of the functional group introduced in the nonwoven fabric N is shown in Table 1.

Table 1

ⁿ

(Example 1)

The non-woven fabric D was cut in a disc with a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 1. Measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 shown below. Results are shown in table 2.

Granulocyte adsorption ratio (%) = {(number of granulocytes in blood before circulation) - (number of granulocytes in blood after circulation)} / (number of granulocytes in blood before circulation) x100 Equation 2

Monocyte adsorption ratio (%) = {(number of monocytes in blood before circulation) - (number of monocytes in blood after circulation)} / (number of monocytes in blood before circulation) x100 Equation 3

Ratio of lymphocyte adsorption (%) = {(number of lymphocytes in blood before circulation) - (number of lymphocytes in blood after circulation)} / (number of lymphocytes in blood before circulation) x100 Equation 4

(Example 2)

The non-woven fabric E was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Example 3)

The non-woven fabric F was cut in a disc having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Example 4)

The nonwoven fabric H was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Example 5)

The non-woven fabric L was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Example 6)

The non-woven fabric M was cut into a disc having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Comparative example 1)

The non-woven fabric C was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Comparative example 2)

The non-woven fabric G was cut into a disc having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Comparative example 3)

The non-woven fabric I was cut into a disc with a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Comparative example 4)

The non-woven fabric J was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Comparative example 5)

The non-woven fabric K was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

(Comparative example 6)

The non-woven fabric N was cut into a disk having a diameter of 8 mm and placed in a polypropylene container. To this vessel, 1 ml of human blood (heparin concentration, 30 U / ml) was added and the tube content was mixed by inversion in an incubator at 37 ° C for 20 minutes. Subsequently, the adsorption ratio of each blood component was calculated. The results are shown in Table 2. The measurement of the number of each blood component was carried out using the XT-1800i Automatic Hematology Analyzer (Sysmex Corporation). The adsorption ratio of each blood component was calculated according to equations 2 to 4 described above. Results are shown in table 2.

[Table 2]

Sample (non-woven fabric) ^of adsorption ^ads of Adsorption ratio of _ocular ocytes m nocytes (%) Example 1 ^{Fabric n} 75.4 _woven 43.8 75 Example 2 ^{Fabric n}

_woven 80.3 45.5 78 Example 3 ^{Fabric n}

_woven 94.1 36.2 95 Example 4 ^{Fabric n} 40.5 _woven 11.4 60 Example 5 ^{Fabric n}

_woven 70.5 40.3 72 Example 6 ^{Fabric n}

_tejida

44,1

0,0

40 Según los resultados de la tabla 2, se descubrió que los vehículos para la adsorción de los componentes sanguíneos de la presente invención en los que un grupo funcional que tiene un grupo funcional ácido se introduce en la superficie del vehículo insoluble en agua muestran relaciones de adsorción notablemente más altas de linfocitos mientras que mantienen las relaciones de adsorción de granulocitos y monocitos, en comparación con los vehículos en los que el grupo funcional en la superficie del vehículo insoluble en agua no tiene un grupo funcional ácido. _woven 77.4 41.2 75 Example Comparative _{woven fabric} 80.0 3.5 85 Example _Woven comparative _fabric 76.3 6.6 90 Example _Woven comparative _{fabric I} 35.2 0.0 26 Example _Woven comparative _{fabric J} 77.7 6.0 91 Example _Woven comparative _fabric 75.2 2.5 85 Example Comparative _fabric n

_woven

44.1

0.0

40 According to the results in Table 2, it was discovered that vehicles for adsorption of blood components of the present invention in which a functional group having an acid functional group is introduced into the surface of the water insoluble vehicle show relationships of Remarkably higher adsorption of lymphocytes while maintaining adsorption ratios of granulocytes and monocytes, compared to vehicles in which the water-insoluble functional group on the surface of the vehicle does not have an acidic functional group.

(Example 7)

The nonwoven fabric D was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of fetal bovine serum (hereinafter referred to as FBS) was prepared so that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml, and the tube contents were mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 then. The results are shown in table 3.

Adsorption ratio of IL-8 (%) = {(concentration of IL-8 before mixing by inversion) - (concentration of IL-8 after mixing by inversion}) / (concentration of IL-8 before investment mix) x 100 Equation 5

Adsorption ratio of IL-17 (%) = {(concentration of IL-17 before mixing by inversion) - (concentration of IL-17 after mixing by inversion}) / (concentration of IL-17 before investment mix) x 100 Equation 6

IFN-y adsorption ratio (%) = {(IFN-y concentration before investment mixing) - (IFN-y concentration after investment mixing}) / (IFN-y concentration before investment mix) x 100 Equation 7

(Example 8)

The non-woven fabric E was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Example 6)

The non-woven fabric F was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Example 7)

The nonwoven fabric H was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Example 8)

The non-woven fabric L was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Example 9)

The non-woven fabric M was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Comparative example 7)

The non-woven fabric C was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Comparative example 8)

The nonwoven fabric G was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared was added so that each of IL-8 is contained at a concentration of 500 pg / ml, and the contents of the tube were mixed by inversion in a 37 ° C incubator during 1 hour. Subsequently, the remaining concentration of IL-8 was measured by ELISA and the adsorption ratio of IL-8 was calculated according to equation 5 above. The results are shown in table 3.

(Comparative example 9)

The nonwoven fabric I was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Comparative example 10)

The non-woven fabric J was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Comparative example 11)

The nonwoven fabric K was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

(Comparative example 12)

The non-woven fabric N was cut into two discs, each with a diameter of 8 mm, and placed in a polypropylene container. To this vessel, 0.8 ml of FBS prepared in such a way that each of IL-8, IL-17 and IFN-y is contained at a concentration of 500 pg / ml was added, and the tube content was mixed by inversion in an incubator at 37 ° C for 1 hour. Subsequently, the remaining concentration of each of IL-8, IL-17 and IFN-y was measured by ELISA, and the adsorption ratios of IL-8, IL-17 and IFN-y were calculated according to equations 5 to 7 previous. The results are shown in table 3.

[Table 3]

^{Relación de adsorción} Relación de adsorción de Relación de adsorción de _{de IL-8 (%} IL-17 (%) IFN-^y (%)Sample (non-woven fabric)

^Adsorption ratio ^Adsorption ratio Adsorption ratio of _{IL-8 (%} IL-17 (%) IFN- ^and (%)

Example 7 97.7 92.5 44.4

Example 8 98.0 90.2 35.3

96,9

95,8

82,1

Example 9

96.9

95.8

82.1

According to the results in Table 3, it was discovered that vehicles for the adsorption of blood components of the present invention in which a functional group having an acid functional group is introduced into the surface of the water insoluble vehicle show adsorption ratios. higher of IL-8, IL-17 and IFN-and compared to vehicles in which the water-insoluble surface functional group does not have an acidic functional group.

INDUSTRIAL APPLICABILITY

The present invention can be used as a column for the adsorption of blood components in the medical care sector.

Claims (6)

1. Vehicle for adsorption of blood components comprising a water insoluble vehicle composed of a fiber or particle, said water insoluble vehicle having a surface on which one or more functional groups are introduced, said one or more functional groups containing a acid functional group selected from the group consisting of the sulfate group, sulphite group and sulfonate group; and that contains an amino group, said fiber having a fiber diameter of or having said particle having a particle diameter of 0.5 to 20 pm, and wherein said water insoluble vehicle has a negative charge amount of 1.0x10-4 to 1.5x10 -3 eq / g. 2. Vehicle for adsorption of blood components according to claim 1, wherein said water insoluble vehicle has a porosity of 85 to 98%. 3. A vehicle for adsorption of blood components according to claim 1 or 2, wherein said water insoluble vehicle has a fiber diameter 4 to 10 pm. 4. A vehicle for the adsorption of blood components according to any one of claims 1 to 3, wherein said acid functional group and said amino group are linked to each other through an alkyl chain. 5. A vehicle for adsorption of blood components according to claim 4, wherein said alkyl chain is an alkyl chain having no more than 3 carbon atoms. 6. Column for adsorption of blood components filled with the vehicle for adsorption of blood components according to any of claims 1 to 5.